Splitomicin S rDNA gene library construction and pyrosequencing S rDNA
2.6.2. 16S rDNA gene library construction and pyrosequencing
2.6.3. 16S rDNA data processing
2.6.4. BioSample accession numbers
All the BioSample sequences are being deposited at the National Center for Biotechnology Information (NCBI; http://ezproxy.student.twu.ca:5115) and will be available under the BioProject ID PRJNA273787 after approval.
2.7. Analytical methods
2.7.1. Gas phase analysis
126.96.36.199. KOH operating procedure
Water containing 9 M KOH in 100 ml gas replacement equipment was used to monitor the carbon dioxide concentration in gas phase. The Splitomicin potential of the KOH solution was regularly measured using gas mixtures containing 0%, 20%, 35%, 80%, and 100% CO2.
188.8.131.52. Gas phase chromatography
Hewlett Packard 5890 Series II gas chromatograph (GC; Agilent Technologies, Santa Clara, CA, USA) equipped with a 30 m long, 0.32 mm id Alltech GAS PRO GSC column (Grace, Deerfield, IL, USA) in series with a 20 m long, 0.25 mm id Chrompack CARBOPLOT P7 column (Agilent Technologies) and a thermal conductivity detector allowed H2, CH4, and CO2 separation and detection. The carrier and reference gas was He for CH4 and CO2 quantification while N2 was used as carrier and reference for H2 analysis. Two gas mixtures were used as standards. First consisted of N2 (15%), CO2 (35%), and CH4 (50%) and the second of H2 (80%) and CO2 (20%). The GC injection port, the thermal conductivity detector chamber, and the oven were respectively maintained at 90, 110, and 55 °C. 25 μl of gas were injected on-column.